海雀稗PvMYB1基因的克隆与表达分析

Cloning and expression analysis of PvMYB1 in Paspalum vaginatum

摘要

为探究MYB转录因子基因是否参与了盐胁迫下对海雀稗（Paspalum vaginatum Sw.）生长发育的调控，本研究分析了盐胁迫对海雀稗生长的影响，克隆了PvMYB1基因，通过生物信息学及基因表达2种分析方法展开研究。结果表明，盐胁迫抑制了海雀稗地上部和根系生长，具体表现为总根长、根表面积及根体积的降低。海雀稗PvMYB1基因全长为1 098 bp，编码365个氨基酸残基，蛋白分子质量为39.26 kDa，属于R2R3-MYB家族，亚细胞定位预测PvMYB1蛋白定位于细胞核中。实时荧光定量PCR结果显示，PvMYB1基因在叶中的表达量高于在根中的表达量；不同时段NaCl处理结果显示，质量浓度为30 g·L⁻¹的NaCl处理显著提升了PvMYB1基因的转录水平。此外，在不同质量浓度的NaCl处理的条件下，当NaCl质量浓度升至15 g·L⁻¹时，根和叶中的PvMYB1基因的表达量显著上调。本研究结果揭示了PvMYB1可能参与了海雀稗对NaCl胁迫的应答。

Abstract

Seashore paspalum (Paspalum vaginatum Sw.) were treated with different concentrations of salt to observe the effect of salt treatment on the growth of seashore paspalum, and the PvMYB1 gene was cloned from seashore paspalum under different treatments and analyzed by using bioinformatics and gene expression analysis methods in a view to investigating whether MYB transcription factor genes are involved in the regulation of growth and development of seashore paspalum under salt stress. The results showed that salt stress inhibited the growth of both shoots and roots in seashore paspalum, manifested by reduction in total root length, root surface area, and root volume. The full length of the seashore paspalumPvMYB1 gene was 1 098 bp, encoding 365 amino acid residues, with a protein molecular weight of 39.26 kDa, and.this gene belongs to the R2R3-MYB family. Subcellular localization prediction indicated that the PvMYB1 protein was located in the nucleus. RT-qPCR analysis demonstrated that the expression of PvMYB1 was higher in leaves than in roots. The results of salt treatment at different time periods showed that 30 g·L^-1 salt treatment significantly increased the transcription level of the PvMYB1 gene. Moreover, under different salt concentration treatments, the expression levels of the PvMYB1 gene in the roots and leaves were significantly upregulated when the salt concentration was 15 g·L^-1. All these results revealed that PvMYB1 is implicated in the response of seashore paspalum to salt stress.

关键词

海雀稗 / 盐胁迫 / 基因表达 / 转录因子

Key words

Paspalum vaginatum / salt stress / gene expression / MYB

参考文献

[1] Balfagón D, Zandalinas S I, Mittler R, et al. High temperatures modify plant responses to abiotic stress conditions[J]. Physiologia Plantarum, 2020, 170(3): 335-344. https://doi.org/10.1111/ppl.13151

[2] Zaman M, Shahid S A, Heng L. Guideline for salinity assessment, mitigation and adaptation using nuclear and related techniques[M]. Cham:Springer, 2018. https://doi.org/10.1007/978-3-319-96190-3

[3] Fao. Global map of salt-affected soils[R]. Rome: FAO, 2021: 1-20.

[4] Liu G Y, Yu X, Zeng Y L, et al. SOS2-FREE1 regulates SOS1 tonoplast sorting to promote Na⁺ compartmentalization in vacuole duringsalt stress response[J]. Journal of Integrative Plant Biology, 2025, 67(10): 2545-2560. https://doi.org/10.1111/jipb.13970

[5] Zhou H P, Shi H F, Yang Y Q, et al. Insights into plant salt stress signaling and tolerance[J]. Journal of Genetics and Genomics, 2024, 51(1): 16-34. https://doi.org/10.1016/j.jgg.2023.08.007

[6] Acosta-Motos J, Ortuño M, Bernal-Vicente A, et al. Plant responses to salt stress: adaptive mechanisms[J]. Agronomy, 2017, 7(1): 18. https://doi.org/10.3390/agronomy7010018

[7] Zhang D W, Zhou H P, Zhang Y, et al. Diverse roles of MYB transcription factors in plants[J]. Journal of Integrative Plant Biology, 2025, 67(3): 539-562. https://doi.org/10.1111/jipb.13869

[8] Li J L, Han G L, Sun C F, et al. Research advances of MYB transcription factors in plant stress resistance and breeding[J]. Plant Signaling & Behavior, 2019, 14(8): 1613131. https://doi.org/10.1080/15592324.2019.1613131

[9] Paz-Ares J, Ghosal D, Wienand U, et al. The regulatory c1 locus of Zea mays encodes a protein with homology to myb proto-oncogene products and with structural similarities to transcriptional activators[J]. The EMBO Journal, 1987, 6(12): 3553-3558. https://doi.org/10.1002/j.1460-2075.1987.tb02684

[10] 李志强, 罗正乾, 徐琳黎, 等. 基于T2T基因组鉴定大豆R2R3-MYB基因家族及干旱和盐胁迫下的表达分析[J]. 生物技术通报, 2025, 41(5): 141-152. https://doi.org/10.13560/j.cnki.biotech.bull.1985.2024-0745

[11] Dubos C, Stracke R, Grotewold E, et al. MYB transcription factors in Arabidopsis[J]. Trends in Plant Science, 2010, 15(10): 573-581. https://doi.org/10.1016/j.tplants.2010.06.005

[12] Li J Y, Liu S Q, Chen P F, et al. Systematic analysis of the R2R3-MYB family in Camellia sinensis: evidence for galloylated catechins biosynthesis regulation[J]. Frontiers in Plant Science, 2022, 12: 782220. https://doi.org/10.3389/fpls.2021.782220

[13] Katiyar A, Smita S, Lenka S K, et al. Genome-wide classification and expression analysis of MYB transcription factor families in rice and Arabidopsis[J]. BMC Genomics, 2012, 13(1): 544. https://doi.org/10.1186/1471-2164-13-544

[14] YOO J H, PARK C Y, KIM J C, et al. Direct interaction of a divergent CaM isoform and the transcription factor, MYB2, enhances salt tolerance in Arabidopsis[J]. Journal of Biological Chemistry, 2005, 280(5): 3697-3706. https://doi.org/10.1074/jbc.m408237200

[15] Yang A, Dai X Y, Zhang W H. A R2R3-type MYB gene, OsMYB2, is involved in salt, cold, and dehydration tolerance in rice[J]. Journal of Experimental Botany, 2012, 63(7): 2541-2556. https://doi.org/10.1093/jxb/err431

[16] Karimi I Y M, Kurup S S, Ali Salem M A M, et al. Evaluation of Bermuda and Paspalum grass types for urban landscapes under saline water irrigation[J]. Journal of Plant Nutrition, 2018, 41(7): 888-902. https://doi.org/10.1080/01904167.2018.1431669

[17] 申晴. 海雀稗种质资源遗传多样性与优异耐盐种质筛选[D]. 海口: 海南大学, 2019. https://doi.org/10.27073/d.cnki.ghadu.2019.001140

[18] 陈静波, 阎君, 姜燕琴, 等. 暖季型草坪草优良选系和品种抗盐性的初步评价[J]. 草业学报, 2009, 18(5): 107-114. https://doi.org/10.3321/j.issn:1004-5759.2009.05.016

[19] 申晴, 韦海燕, 卞华, 等. 海雀稗种质资源的耐盐性评价[J]. 热带生物学报, 2020, 11(1): 11-19. https://doi.org/10.15886/j.cnki.rdswxb.2020.01.003

[20] 黄杰, 宋剑灵, 安娜, 等. 柱花草磷饥饿响应基因SgPHR1和SgPHR2的克隆与表达分析[J]. 热带作物学报, 2021, 42(8): 2158-2166. https://doi.org/10.3969/j.issn.1000-2561.2021.08.006

[21] 邹晓燕, 王林杰, 黄杰, 等. 柱花草SgNramp1和SgNramp2基因克隆与表达分析[J]. 草地学报, 2022, 30(6): 1388-1395. https://doi.org/10.11733/j.issn.1007-0435.2022.06.009

[22] 刘莉婷, 高佳淼, 周鸿明, 等. 地毯草扩张蛋白基因AcEXPA1的克隆与表达分析[J]. 草地学报, 2025, 33(2): 351-359. https://doi.org/10.11733/j.issn.1007-0435.2025.02.003

[23] 张郎织, 李季肤, 黄杰, 等. 地毯草AcMATE1基因的克隆与表达分析[J]. 热带作物学报, 2021, 42(7): 1860-1867. https://doi.org/10.3969/j.issn.1000-2561.2021.07.007

[24] Park H J, Kim W Y, Yun D J. A new insight of salt stress signaling in plant[J]. Molecules and Cells, 2016, 39(6): 447-459. https://doi.org/10.14348/molcells.2016.0083

[25] Campanelli A, Ruta C, Morone-Fortunato I, et al. Alfalfa (Medicago sativa L.) clones tolerant to salt stress: in vitro selection[J]. Open Life Sciences, 2013, 8(8): 765-776. https://doi.org/10.2478/s11535-013-0194-1

[26] Marcum K B, Murdoch C L. Salinity tolerance mechanisms of Six C₄ turfgrasses[J]. Journal of the American Society for Horticultural Science, 1994, 119(4): 779-784. https://doi.org/10.21273/jashs.119.4.779

[27] Liao Y, Zou H F, Wang H W, et al. Soybean GmMYB76, GmMYB92, and GmMYB177 genes confer stress tolerance in transgenic Arabidopsis plants[J]. Cell Research, 2008, 18(10): 1047-1060. https://doi.org/10.1038/cr.2008.280

[28] 陈娜, 迟晓元, 潘丽娟, 等. MYB转录因子在植物盐胁迫调控中的研究进展[J]. 植物生理学报, 2015, 51(9): 1395-1399. https://doi.org/10.13592/j.cnki.ppj.2015.0239

[29] 王文林, 陈海生, 郑树芳, 等. 澳洲坚果MiMYB2基因克隆及结构与功能分析[J]. 植物研究, 2020, 40(6): 913-922. https://doi.org/10.7525/j.issn.1673-5102.2020.06.014

[30] Sairam R, Tyagi A. Physiology and molecular biology of salinity stress tolerance in plants[J]. Current Science, 2004, 86(3): 407-421.

[31] Zhao Y, Tian X J, Wang F, et al. Characterization of wheat MYB genes responsive to high temperatures[J]. BMC Plant Biology, 2017, 17(1): 208. https://doi.org/10.1186/s12870-017-1158-4

[32] Ren Z Z, Zhang P Y, Su H H, et al. Regulatory mechanisms used by ZmMYB39 to enhance drought tolerance in maize (Zea mays) seedlings[J]. Plant Physiology and Biochemistry, 2024, 211: 108696. https://doi.org/10.1016/j.plaphy.2024.108696

[33] Liu L, Li H Y, Wang X Y, et al. Transcription factor TaMYB30 activates wheat wax biosynthesis[J]. International Journal of Molecular Sciences, 2023, 24(12): 10235. https://doi.org/10.3390/ijms241210235

[34] Li W H, Zhong J L, Zhang L H, et al. Overexpression of a Fragaria vesca MYB transcription factor gene (FvMYB82) increases salt and cold tolerance in Arabidopsis thaliana[J]. International Journal of Molecular Sciences, 2022, 23(18): 10538. https://doi.org/10.3390/ijms231810538

[35] Tang Y H, Bao X X, Zhi Y L, et al. Overexpression of a MYB family gene, OsMYB6, increases drought and salinity stress tolerance in transgenic rice[J]. Frontiers in Plant Science, 2019, 10: 168. https://doi.org/10.3389/fpls.2019.00168

[36] Du B Y, Liu H, Dong K T, et al. Over-expression of an R2R3 MYB gene, MdMYB108L, enhances tolerance to salt stress in transgenic plants[J]. International Journal of Molecular Sciences, 2022, 23(16): 9428. https://doi.org/10.18488/journal.105

[37] Zhang L C, Zhao G Y, Jia J Z, et al. Molecular characterization of 60 isolated wheat MYB genes and analysis of their expression during abiotic stress[J]. Journal of Experimental Botany, 2012, 63(1): 203-214. https://doi.org/10.1093/jxb/err264

[38] Gong Q Y, Li S, Zheng Y, et al. SUMOylation of MYB30 enhances salt tolerance by elevating alternative respiration via transcriptionally upregulating AOX1a in Arabidopsis[J]. The Plant Journal, 2020, 102(6): 1157-1171. https://doi.org/10.1111/tpj.14689

[39] Zhang L Y, Jiang X C, Liu Q Y, et al. The HY5 and MYB15 transcription factors positively regulate cold tolerance in tomato via the CBF pathway[J]. Plant, Cell & Environment, 2020, 43(11): 2712-2726. https://doi.org/10.1111/pce.13868

基金资助

国家自然科学基金项目(32060409)

国家自然科学基金项目(32371782)

国家自然科学基金项目(32460358)

海南省科技人才创新基金项目(KJRC2023C21)

海南大学南繁与热带高效农业研究协同创新中心项目(XTCX2022NYB08)

海南大学生态文明协同创新中心项目(XTCX2022STC10)